Microalgae are a diverse group of organisms with significant potential for industrial applications: as feedstock in aquaculture as well as in the production of valuable bioproducts such as lipids, carotenoids and enzymes. Lately, developments in molecular biology have improved production yields of algae bioproducts, thus increasing their industrial relevance. Additionally, variations in bioprocessing factors (i.e. temperature, pH, light, carbon source, salinity, nutrients, etc.) have been used to enhance both biomass and specific bioproducts' productivities. Particularly, microalgae have increasingly gained research interest as a source of specialty lipids such as arachidonic, eicosapentaenoic and docosahexaenoic acids, which are often reported in literature to provide several health benefits. Moreover, there has been a recent resurgence in interest in microalgae as an oil producer for biofuel applications. Significant advances have also been made in upstream processing to generate cellular biomass and oil. However, extracting and purifying oil from algae continues to prove a significant challenge in producing both microalgae bioproducts and biofuel, as microbial oil extraction is relatively energy-intensive and costly. Thus, developing inexpensive and robust oil extraction and purification processes is a major challenge facing both the microalgae to bioproduct, and biofuel industries. This paper presents an overview, based on the last 10 years, of advances made in technologies for extracting and purifying microalgae oil. We compared solvent extraction technologies with extraction alternatives such as mechanical milling and pressing, enzymatic and supercritical fluid extraction. We also reviewed recent advances based on molecular engineering of microbes to aid oil extraction. Downstream processing for the potential commercial production of microalgae oil not only must consider economic costs, but should also consider minimizing environmental impacts in order to attain sustainable production processes.
Omega-3 fatty acids, namely docosahexaenoic acid and eicosapentaenoic acid, have been linked to several beneficial health effects (i.e. mitigation effects of hypertension,
Various extraction methods were assessed in their capacity to extract fatty acids from a dried biomass of Thraustochytrium sp. ONC-T18. Direct saponification using KOH in ethanol or in hexane:ethanol was one of the most efficient techniques to extract lipids (697 mg g(-1)). The highest amount of fatty acids (714 mg g(-1)) was extracted using a miniaturized Bligh and Dyer extraction technique. The use of ultrasonics to break down cell walls while extracting with solvents (methanol:chloroform) also offered high extraction yields of fatty acids (609 mg g(-1)). Moreover, when the transesterification mixture used for a direct transesterification method was doubled, the extraction of fatty acids increased approximately 77% (from 392 to 696 mg g(-1)). This work showed that Thraustochytrium sp. ONC-T18 has the ability to produce over 700 mg g(-1) of lipids, including more than 165 mg g(-1) of docosahexaenoic acid, which makes this microorganism a potential candidate for the commercial production of polyunsaturated fatty acids. Finally, other lipids, such as myristic, palmitic, palmitoleic, and oleic acids, were also produced and recovered in significant amounts (54, 196, 123, and 81 mg g(-1)), respectively.
To the best of our knowledge, stability studies on astaxanthin contained in carotenoproteins extracted from lactic acid fermented shrimp byproduct have never been reported. Carotenoprotein powder, containing 1% free astaxanthin, was subjected to oxidation factors of illumination, oxygen availability, and temperature, using synthetic astaxanthin as a control. The individual effects as well as first and second degree interactions were studied on natural and synthetic free astaxanthin stability. Air and full light were the two individual factors with the highest effects on astaxanthin oxidation. Sixty-two and 46% natural and synthetic astaxanthin, respectively, oxidized when exposed to air for 8 weeks of storage, whereas 35 and 28% of natural and synthetic astaxanthin, respectively, oxidized under full light. Ninety-seven and 88% of natural and synthetic astaxanthin, respectively, oxidized under a combination of full light, air, and 45 degrees C at 8 weeks of storage. Storage in the dark, nonoxygen, and 25 degrees C were the treatments that efficiently minimized astaxanthin oxidation. Natural astaxanthin from fermented shrimp byproduct presented moderate stability levels. Although natural astaxanthin oxidized faster than the synthetic pigment, its stability may improve by antioxidant and polymer addition.
Astaxanthin (3,3'-dihydroxy-4,4'-diketo-β-carotene) (1) is a carotenoid of significant commercial value due to its superior antioxidant potential, application as a component of animal feeds, and ongoing research that links its application to the treatment and prevention of human pathologies. The high commercial cost of 1 is also based upon its complex synthesis. Chemical synthesis has been demonstrated, but produces a mixture of stereoisomers with limited applications. Production from biological sources is limited to natural producers with complex culture requirements. The biosynthetic pathway for 1 is well studied; however, questions remain that prevent optimized production in heterologous systems. Presented is a direct comparison of 12 β-carotene (2) hydroxylases derived from archaea, bacteria, cyanobacteria, and plants. Expression in Escherichia coli enables a comparison of catalytic activity with respect to zeaxanthin (3) and 1 biosynthesis. The most suitable β-carotene hydroxylases were subsequently expressed from an efficient dual expression vector, enabling 1 biosynthesis at levels up to 84% of total carotenoids. This supports efficient 1 biosynthesis by balanced expression of β-carotene ketolase and β-carotene hydroxylase genes. Moreover, our work suggests that the most efficient route for astaxanthin biosynthesis proceeds by hydroxylation of β-carotene to zeaxanthin, followed by ketolation.
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